Pathological Damage Research Articles

Twist2 knockdown alleviates renal ischemia-reperfusion injury by maintaining mitochondrial function and enhancing mitophagy through Bnip3.

Renal ischemia-reperfusion injury (IRI) is a major cause of acute kidney injury (AKI). Twist-related protein 2 (Twist2) is a basic helix/loop/helix transcription factor. However, the underlying effects of Twist2 in IRI remain to be elucidated. Herein, we found that the expression of Twist2 was significantly upregulated in renal tissues of mice induced by ischemia/reperfusion (I/R) and in human renal tubular epithelial cell HK-2 exposed to hypoxia-reoxygenation. We silenced Twist2 with RNAi technology. Twist2 knockdown alleviated renal pathological damage in mice. Twist2 depletion ameliorated IRI-induced mitochondrial dysfunction, such as increasing ATP content and mitochondrial DNA copy number and restoring mitochondrial membrane potential in the kidneys of mice, and similar results were observed in in vitro experiments. Twist2 interference increased the expression of LC3B and decreased the expression of p62 and mitochondrial membrane proteins TIMM23 and TOMM20 both in vivo and in vitro. Electron microscope and the co-localization of LC3B and mitotracker DsRed suggested the induction of autophagy and mitophagy after Twist2 knockdown in kidneys or cells. Mechanism studies revealed that Twist2 exerted a direct inhibitory effect on BCL2 interacting protein 3 (Bnip3) transcriptional activity by targeting the Bnip3 promoter. In hypoxia/reoxygenation-induced renal tubular epithelial cells, the interference of Bnip3 reversed the effect of Twist2 depletion on mitochondrial function and mitophagy. In conclusion, our findings suggest that the depletion of Twist2 exerts renoprotective effect in I/R-induced AKI. Twist2 regulates mitochondrial function and mitophagy in part by targeting and downregulating Bnip3. Our study provides new insights into the pathological mechanisms of I/R-induced AKI.

Investigation of the role and mechanism of dapagliflozin in mitigating renal injury in rats afflicted with diabetic kidney disease.

The etiology of diabetic kidney disease (DKD) is multifaceted, with hyperglycemia, inflammation, oxidative stress, and fibrosis recognized as key contributors to renal damage in individuals with DKD. Clinical evidence suggests that dapagliflozin not only reduces blood glucose levels but also demonstrates superior efficacy in ameliorating pancreatic islet cell injury while preserving cardiac and renal function. However, the precise underlying mechanism has been poorly elucidated in the current literature. In this study, a DKD rat model was established by administering a single intraperitoneal injection of streptozotocin (STZ) to investigate the renoprotective properties of dapagliflozin and its underlying mechanisms. The findings of this study indicate that dapagliflozin enhanced pancreatic islet cell function, lowered blood glucose levels, and significantly reduced biochemical markers and renal pathological damage in DKD rats. Dapagliflozin also exerted anti-inflammatory, antioxidant, and antifibrotic effects by inhibiting the activation of the p38 MAPK/NF-κB pathway, enhancing the activity of the SIRT1/Akt/GSK-3β/Nrf2/HO-1 signaling pathway, and inhibiting the over-activation of the TGF-β1/Smad2/3 signaling pathway. These effects led to a reduction in renal injury and improved renal function in DKD rats.

Yueju Pill Inhibits Apoptosis by Regulating the SCF/c-Kit/PI3K/AKT Signaling Pathway to Ameliorate Functional Dyspepsia.

This study aimed to investigate the therapeutic effect of Yueju Pill (YJP) on functional dyspepsia (FD) rats and its mechanism of promoting gastrointestinal motility. We replicated FD rat models using classical tail pinching and irregular feeding for 14 days. After 28 days of YJP treatment, we measured the gastric emptying rate and intestinal propulsion rate of the rats. Hematoxylin-eosin (H&E) staining was used to observe the pathological damage in the gastric antrum. The serum levels of related brain-gut peptides (BGPs) were determined using the enzyme-linked immunosorbent assay (ELISA). Furthermore, we detected the expression of proteins related to the SCF/c-Kit/PI3K/AKT signaling pathway through Western blot and immunohistochemistry. Finally, we assessed the levels of apoptosis using the TUNEL assay. YJP improved gastric emptying and small intestine propulsion rates while reducing gastric tissue injury in FD rats. Moreover, YJP increased the levels of gastrin (GAS) and ghrelin (Ghrelin) and decreased the levels of cholecystokinin (CCK) and vasoactive intestinal peptide (VIP). YJP also elevated the levels of SCF, c-kit and Bcl-2, promoted the phosphorylation of PI3K and AKT, and inhibited the expression of Bax. YJP achieved the effect of FD treatment by regulating the SCF/c-Kit/PI3K/AKT pathway, providing a theoretical basis for the clinical treatment of FD.

The immunogenicity of PRV ΔgE/TK/UL49.5 three-gene-deleted vaccine in mice

BackgroundPseudorabies (PR) caused by the re-emerging of pseudorabies virus (PRV) variant has outbroken among PRV vaccine immunized swine in many pig farms, which has caused serious social and economic consequences since the end of 2011. The PRV UL49.5 protein can inactivate the transporter associated with antigen processing (TAP), thereby downregulating the cell surface expression of swine leukocyte antigen class I (SLA-I) to evade host immune surveillance.MethodsIn this study, based on the PRV ΔgE/TK strain, PRV ΔgE/TK/UL49.5 triple gene deletion strain was constructed through homologous recombination and deletion of the PRV UL49.5 gene by the Cre-LoxP system. Its growth curve and effect on SLA-I transcription level were determined. Preliminary studies were carried out on serum neutralizing antibody levels, IFN-γ and IL-4 cytokines levels in mice immunized with PRV ΔgE/TK/UL49.5, and the viral load and challenge protection in mice tissues after challenge.ResultsThe growth characteristics of PRV ΔgE/TK/UL49.5 strain were similar to those of PRV ΔgE/TK strain. The level of SLA-I was returned to normal after the deletion of PRV UL49.5 gene. The immunization of PRV ΔgE/TK/UL49.5 did not affect the weight gain of mice. Immunized mice could induce high levels of serum neutralization antibodies and immune cytokines, including IFN-γ and IL-4, which could provide complete protection against virulent PRV challenge. No obvious pathological damage was observed in lung, brain and trigeminal ganglion of mice immunized with PRV ΔgE/TK/UL49.5, and the tissue viral load was the lowest.ConclusionsPRV ΔgE/TK/UL49.5 strain can induce enhanced immunogenicity and had the potential to be used as a candidate strain.

Establishment of an animal model for monkeypox virus infection in dormice

This study aims to establish an animal model of monkeypox virus (MPXV) infection in dormice through intranasal inoculation. Male dormice aged 4–5 months were selected as experimental subjects and administered different titers of MPXV (103.5 PFU, 104.5 PFU, and 105.5 PFU, respectively) via nasal instillation. Within 14 days post-infection, clinical indicators such as survival rate, body weight changes, respiratory status, and mental state were continuously monitored. Additionally, tissue samples from the lungs, liver, spleen, and trachea of dormice from each group were collected on the 5th and 10th days for virus titer detection, and histopathological analysis was performed on lung samples collected on the 5th and 10th days. Dormice infected with MPXV exhibited typical symptoms such as appetite loss, continuous body weight reduction, aggravated respiratory difficulties, accompanied by lethargy, chills, and other clinical manifestations similar to human monkeypox infection. Virological tests further confirmed the distribution of MPXV in multiple vital organs of dormice, including the lungs, liver, spleen, and trachea, with particularly significant pathological damage observed in lung tissue. An MPXV infection model in dormice was successfully established through intranasal inoculation with a titer of 105.5 PFU MPXV, which can be used for studying the infection mechanism and pharmacology of MPXV.

Administration of a Recombinant Fusion Protein of IFN-γ and CD154 Inhibited the Infection of Chicks with Salmonella enterica

The cytokines IFN-γ and CD154 have been well established, and they play pivotal roles in immune protection against Salmonella in mice, but their effects and specific mechanisms in Salmonella-infected chickens are less understood. In this study, we conducted animal experiments to screen the highly immunoprotective chIFN-γ-chCD154 fusion protein compared with single protein chIFN-γ or chCD154 in white Leghorn chickens. The results showed that compared with separate pretreatments with chIFN-γ and chCD154, the fusion protein, chIFN-γ-chCD154, synergistically increased survival of infected chickens, reduced bacterial load in feces and organs, and attenuated pathological damage to the liver and cecum. Pretreatment with chIFN-γ-chCD154 also increased humoral immune responses, expression of the tight junction proteins zo-1, occludin, and claudin-1, and the relative abundance of Enterococcus_cecorum, Lactobacillus_helveticus, and Lactobacillus_agilis, which protect against intestinal inflammation. Compared with single protein pretreatment, chIFN-γ-chCD154 significantly upregulated STAT1, IRF1, and GBP1 in infected chickens while decreasing mRNA expression of TLR4, MyD88, NF-κB, TNF-α, IL-6, and IL-1β. In summary, damage to the cecal epithelial barrier and the inflammation induced by S. typhimurium infection was alleviated by chIFN-γ-chCD154 pretreatment through a mechanism involving the TLR4/MyD88/NF-κB and IFN-γ/STAT/IRF1/GBP1 pathways.

The mechanism of Bovis Culus Sativus protecting BBB damage in stroke: Insights from network pharmacology, bioinformatics, and experiments.

Bovis calculus (BC) has a medicinal history of over 2000 years in treating stroke in China. Bovis Culus Sativus (BCS) has similar pharmacological effects to BC. Due to the scarcity of BC, BCS is often used as a substitute for BC in clinical practice for treating stroke in traditional Chinese medicine. This study aims to investigate the inhibitory effect of BCS on blood-brain barrier (BBB) damage following stroke, and to elucidate the molecular basis of BCS neuroprotection through network pharmacology and bioinformatics. The contents of bilirubin and bile acids in BCS were quantified using HPLC. A cerebral ischemia-reperfusion injury (CIRI) rat model was established to assess neurological function, cerebral infarction, pathological damage, and Evans Blue staining. R language was used to analyze GEO public data to identify therapeutic targets for ischemic stroke. Public databases and literature were utilized to screen for active components of BCS, and the Swiss Target Prediction database was used to predict the active drug targets. Network pharmacology analysis was conducted on drug and disease targets, followed by immune infiltration and molecular docking of key targets. Finally, ELISA, RT-PCR, Western blot, IHC, and TEM were employed to validate the effectiveness of the targets. The content of bile acids and bilirubin in the tested BCS was 6.9% and 37.89%, respectively. The study showed that BCS reduced neurological function scores and cerebral infarction rates in stroke rats, prevented Evans Blue leakage, and mitigated histopathological damage in the ischemic brain region. Additionally, BCS improved the structural and functional integrity of the BBB, enhancing the expression of Occludin, ZO-1, and Claudin-5 while downregulating the expression of MDR1, aquaporin-4, MMP-9, and MMP2. Bioinformatics and network pharmacology analyses indicated that the therapeutic effects of BCS in stroke are primarily associated with the inhibition of inflammatory pathways, including TNF, NFKB, and MAPK. ELISA, RT-PCR, and Western blot results further confirmed that BCS significantly suppressed neuroinflammation in stroke rats. BCS shows promising efficacy against ischemic stroke, maintaining the function and structural integrity of the BBB. Its protective effect on the BBB may be related to the inhibition of the TNF-NFκB-MAPK signaling pathways.

In Situ Dual-targeted Drug Delivery System for Alleviating Imaging and Pathological Damage in Septic Arthritis.

Septic arthritis is a severe disease that damages articular cartilage and triggers a strong inflammatory response. Current treatments mainly depend on systemic antibiotics and lack effective intra-articular therapies, as well as standardized animal models, and precise detection methods. In this study, we present a drug delivery system responsive to the bacterial microenvironment for targeted inflammation control, along with an effective method for monitoring changes in septic arthritis in SD rats. This system consists a core with pH-sensitive metal-organic frameworks ZIF-8 loading anti-inflammatory drugs indomethacin and a shell with hybrid cell membranes from macrophages (MM) and platelets (PM), refer as MP@ZIF-8@IN. This system, which diverges from traditional treatments, enhances drug utilization, prolongs local retention, and allows for spontaneous release at the treatment site, thereby enabling the exclusive intra-articular treatment of septic arthritis. The drug delivery system inhibits the NF-κB pathway, reduces oxidative stress, and regulates macrophage polarization, preventing cartilage destruction. Additionally, in this standardized animal model utilizing the knee joints of SD rats, we have developed musculoskeletal ultrasound and magnetic resonance imaging for time-based monitoring, thus overcoming the limitation of conventional methods, which are unsuitable for soft tissue analysis. Our findings advance therapeutic strategies for septic arthritis and encourage further application of visualization techniques in related fields. STATEMENT OF SIGNIFICANCE: This study presents significant advancements in the treatment and understanding of septic arthritis. Our customized drug delivery system targets bacteria and macrophages, ensuring long-time drug retention and enhanced inflammation control, all while reducing reliance on antibiotics-an important step toward addressing antibiotic resistance. Additionally, we have refined septic arthritis animal models to establish clearer guidelines for intervention timing, grounded in clinical symptoms and imaging data. This addresses a critical gap in current research and offers a practical framework for future therapeutic approaches.

Investigation on bowel regulation and constipation relief based on the microbial fermentation solution of cassia seed.

As digestive health issues rise and interest in natural therapies grows, traditional herbs like Cassia Seed are gaining attention for their antioxidant, laxative, and digestive benefits. This study aimed to optimize the fermentation conditions of Cassia seed using microbial technology to enhance the content of anthraquinone compounds, thereby augmenting its pharmacological effects, particularly in promoting intestinal peristalsis and alleviating constipation. Fermentation of Cassia Seed was conducted under controlled microbial conditions. High-performance liquid chromatography (HPLC) was employed to analyze the anthraquinone profile of the fermented solution. Mouse model was utilized to evaluate biochemical markers and intestinal functions, while in the zebrafish constipation model, calcein was utilized as a marker to monitor intestinal function and excretion dynamics. Cassia Seed was fermented under controlled microbial conditions. HPLC was employed to analyze the anthraquinone profile of the fermented solution. Mouse models were utilized to evaluate biochemical markers and intestinal functions, while a zebrafish constipation model used calcein as a marker to monitor intestinal function and excretion dynamics. The optimized fermentation significantly increased the content of anthraquinone compounds, including substances like emodin, aloe-emodin, rhein, chrysophanol, and physcion, as confirmed by HPLC analysis. In a mouse constipation model, the fermented solution reduced nitric oxide (NO) levels while increasing acetylcholine (Ach) and gastrin (Gas) levels, thereby promoting intestinal peristalsis and accelerating bowel movements. Additionally, the fermented solution facilitated repair of the colonic mucosa in mice without significant pathological damage such as degeneration or fibrous tissue proliferation. In the zebrafish model, the fermented Cassia Seed solution reduced calcein fluorescence intensity, indicating enhanced intestinal peristalsis and accelerated bowel movements, thereby decreasing calcein retention time in the body. This study investigated the potential clinical applications of fermented Cassia seed ferment in digestive health, providing a theoretical basis and inspiration for further research and development. Additionally, it offers new insights and possibilities for the development of novel natural medicines or functional foods aimed at improving constipation and promoting intestinal health.

A vaccine combining ORF132 and ORF25 expressed by Saccharomyces cerevisiae induces protective immunity in Carassius auratus gibelio against CyHV-2.

CyHV-2 is the pathogen of herpesvirus haematopoietic necrosis (HVHN), resulting in significant economic losses in crucian carp. Although multiple oral vaccines have been developed to prevent CyHV-2, they have not achieved ideal protective effects. To improve the protective effect of oral vaccine, a combination vaccine was conducted by mixing recombinant Saccharomyces cerevisiae displaying ORF132 or ORF25 on the cell surface in a 1:1 ratio. Oral immunization with this combined vaccine induced specific antibodies against ORF132 and ORF25 in fish serum, as well as increased the transcription levels of IgT, IgM, IL-1β, IFN-1, and TNF-α in the spleen, head-kidney, and hindgut. Importantly, the combined vaccine provided significant protection against CyHV-2 infection in crucian carp, resulting in a relative percent survival of 80.77%. Additionally, it suppressed viral load and alleviated pathological damage of CyHV-2-infected fish. The results suggest that the combined vaccine exhibits a better protective effect compared to the single-antigen vaccines, indicating it as a promising approach to prevent CyHV-2 outbreaks.

Multi-omics elucidates the kidney damage caused by aquatic Cu via the gut-kidney axis in ducks.

Copper (Cu) is an essential trace element for biological growth and development. Excessive intake of Cu exists harmful effects on organisms. However, whether excessive Cu intake induces kidney function damage by gut microbiota regulation remains unclear. Ducks are important species of waterfowl that are often exposed to Cu contamination in water sources. In this study, we aim to elucidate the effects of Cu exposure on renal inflammation through the gut-kidney axis in ducks. The ducks were gavaged with different doses of CuSO4 (0, 100, and 200 mg/kg body weight) for 4 weeks. Results indicate that Cu exposure causes pathological damage to the kidney, with a significant increase in the levels of TNFα, IL-6, and IL-1β in both serum and renal tissue. 16S rDNA analysis revealed that the relative abundances of Candidatus_Saccharimonas and Bacteroides were significantly reduced in the Cu-induced group. Transcriptomic analysis of kidney tissue reveals that following Cu exposure, 30 genes show significant differential expression. GO and KEGG enrichment analyses were most involved in Interleukin-1 Receptor Activity, Taurine and hypotaurine metabolism, Nitrogen metabolism, and Proximal tubule bicarbonate reclamation. Metabolomic analysis revealed that 28 metabolites are present in both kidney tissue and cecal contents. Correlation analysis revealed a strong correlation among 5 common metabolites: Aminoglutethimide, Boscalid, Dantrolene, Cer[ns] d34:1, and Stearidonic acid. In the cecum, these five metabolites are closely associated with 26 intestinal microorganisms, including Bacteroides, Candidatus_Saccharimonas, and Colidextribacter. In the kidney, apart from Stearidonic acid, the other four metabolites are closely correlated with genes such as FOS, and IL1RL1. Overall, our study indicates that excessive Cu induces significant kidney inflammation, the metabolites alteration and gut microbiota disorders. These findings shed light on the underlying mechanisms of Cu-induced kidney damage via the indirect pathway of the gut-kidney axis.

Fe2+ inhibits spring viraemia of carp virus proliferation via ATG14-dependent autophagy

Iron (Fe) plays an essential role in diverse cellular activities, yet its participation in the context of fish virus infection remains elusive. In this work, we present findings on the antiviral efficacy of Fe2+ in instigating autophagic clearance of spring viraemia of carp virus (SVCV). In vivo, Fe2+ increases the survival rate of zebrafish infected with SVCV, mitigates pathological damage, and diminishes viral load in liver tissues. In vitro, Fe2+ impedes the proliferation of SVCV in cells. Further investigations reveal that Fe2+ stimulates cellular autophagy and induce autophagy post-viral infection, with the antiviral function of Fe2+ being compromised upon autophagy activation blockade. Mechanistically, the Fe2+-mediated clearance of SVCV virus relies on the pivotal autophagic initiation factor, autophagy related 14 (ATG14). This study elucidates the mechanism through which Fe2+ induces autophagic for viral clearance, underscoring the potential of low concentrations of exogenous iron as an efficacious therapeutic strategy for fish viral diseases.

Ophiopogonin D Alleviates Sepsis-Induced Acute Lung Injury Through Improving Microvascular Endothelial Barrier Dysfunction via Inhibition of HIF-1α-VEGF Pathway.

Pulmonary endothelial barrier dysfunction is a hallmark of sepsis-induced acute lung injury (ALI). Ophiopogonin D (OP-D), isolated from the roots of Ophiopogon japonicus, is involved in regulating inflammation, apoptosis and intestinal permeability. However, the role of OP-D in ALI has not been reported and the related mechanisms remain unclear. In this study, cecal ligation and puncture (CLP) was used to establish a septic ALI model in mice. We found that OP-D effectively alleviated lung pathological damage. Moreover, OP-D decreased pulmonary microvascular permeability, restrained the inflammatory response and apoptosis in murine lung tissues and LPS-exposed PMVECs. Specifically, OP-D exerted the beneficial effects via mediating the inactivation of HIF-1α-VEGF pathway, which was partly abrogated by the overexpression of HIF-1α. Collectively, our findings showed that OP-D protected against sepsis-induced ALI through improving pulmonary microvascular endothelial barrier dysfunction via suppressing HIF-1α-VEGF pathway.

Xuefu Zhuyu Decoction improves hyperlipidemia through the MAPK/NF-κB and MAPK/PPARα/CPT-1A signaling pathway.

Xuefu Zhuyu Decoction (XZD) is widely used in the treatment of cardiovascular diseases. The purpose of this study was to explore the pharmacological effects and molecular mechanisms of XZD in improving hyperlipidemia and to provide a theoretical framework for clinical application. In this study, the signaling pathways regulated by XZD in improving hyperlipidemia were predicted by network pharmacology. Molecular docking was used to verify the affinity between the components in XZD and the target. Furthermore, a hyperlipidemic model in rats was constructed through feeding a high-fat diet. The effect of XZD on hyperlipidemia was verified by histopathological staining, Elisa, and western blot. The results found that the XZD improved dyslipidemia and inflammatory factor disorders, and inhibited liver function damage, pathological damage, and oxidative stress damage in hyperlipidemic rats. The findings from molecular docking and network pharmacology suggested that the mechanism of XZD improving hyperlipidemia may be closely related to the MAPK, NF-κB, and PPAR pathways. This study demonstrated that the XZD inhibited liver lipid metabolism disorder and inflammatory response by regulating the MAPK/NF-κB and MAPK/PPARα/CPT-1A pathway, significantly improved liver histopathological damage and oxidative stress injury, and played a protective role in hyperlipidemic rats.

Rescue of CUMS-induced HPA axis hyperfunction and hypothalamic synaptic deficits by Citrus aurantium L. cv. Daidai essential oil via the cAMP/PKA/Grin2b pathway.

Traditional Chinese medicine has historically used Citrus aurantium L. cv. Daidai to alleviate anxiety and improve mood. Essential oils are the primary active component of Citrus aurantium L. cv. Daidai, but little research has been conducted on the active substances and mechanisms of the antidepressant effect of Citrus aurantium L. cv. Daidai essential oil (CEO). This research used network pharmacology, molecular docking, transcriptomics, and in vitro and in vivo experimental validation to assess the effectiveness and therapeutic mechanism of CEO. We used gas chromatography‒mass spectrometry(GC-MS) to identify and quantify CEO components and brain-penetrating chemicals. CEO was given to chronic and unpredictable mild stimulation (CUMS) mice for 4 weeks. Depression was assessed using sucrose preference, forced swimming, tail suspension, elevated plus-maze, and open field tests. Analyzing brain homogenates and serum biochemistry data simultaneously revealed neurotransmitter and hormone alterations. Tissue samples were collected for histological and protein analysis. Furthermore, transcriptome and network pharmacology were used to investigate the mechanism by which CEO alleviates depression, and in vitro glutamate(Glu)-induced PC12 cell injury assays were conducted to validate this new mechanism. CEO inhalation altered neurotransmitter and hormone expression and improved CUMS-induced weight and depression-like behavior in mice. Compared with CUMS mice, CEO mice presented less pathological brain damage, as demonstrated by HE staining, immunohistochemistry, Golgi staining, transmission electron microscopy, and immunofluorescence staining. We discovered that 13 of the active chemicals in CEO act on 522 targets, 96 of which are linked to depression. PRKACA was identified as the core target by a modular analysis of the PPI network. Network pharmacology and transcriptomics revealed that CEO influences depression via the cAMP/PKA/Grin2b pathway and Glu synaptic modulation. In vivo studies indicated that CEO administration reduced PKA and Grin2b phosphorylation in CUMS mice, inhibited the cAMP/PKA/Grin2b pathway, protected against synaptic deficits, and restored HPA axis function. In vitro investigations revealed that the survival rate of PC12 cells treated with CEO increased, the apoptotic rate decreased, and the expression of LDH, Ca2+, and MDA decreased. Western blot and immunofluorescence staining indicated that CEO inhibits the cAMP/PKA/Grin2b pathway to regulate Glu-induced PC12 cells and that 8-Bromo-cAMP pretreatment reduces the protective effect of CEO. The active chemicals in CEO can inhibit the cAMP/PKA/Grin2b pathway, reduce anxiety and depression, alleviate excitotoxicity caused by Glu synaptic overactivation, protect against hypothalamic synaptic deficits, and restore HPA axis function. This research could serve as a reference for the field of illness prevention and complementary therapies for depression.

Exosomal miR-122 derived from M2 macrophages induces osteogenic differentiation of bone marrow mesenchymal stem cells in the treatment of alcoholic osteonecrosis of the femoral head

Alcoholic osteonecrosis of the femoral head (AIONFH) is caused by long-term heavy drinking, which leads to abnormal alcohol and lipid metabolism, resulting in femoral head tissue damage, and then pathological necrosis of femoral head tissue. If not treated in time in clinical practice, it will seriously affect the quality of life of patients and even require hip replacement to treat alcoholic femoral head necrosis. This study will confirm whether M2 macrophage exosome (M2-Exo) miR-122 mediates alcohol-induced BMSCs osteogenic differentiation, ultimately leading to the inhibition of femoral head necrosis. M2 macrophages were identified by flow cytometry, and the isolated exosomes were characterized by transmission electron microscopy (TEM) and Nanoparticle Tracking Analysis (NTA). Next, miR-122 was overexpressed by transfecting miR-122 mimic, and the expression of miR-122 in M2 macrophages and their exosomes was evaluated. Subsequently, the effect of exosomal miR-122 on the osteogenic differentiation ability of BMSCs was detected, including cell proliferation, expression of osteogenic-related genes (RUNX2, BMP2, OPN, ALP), and calcium nodule formation. Finally, the therapeutic effect of M2-Exo was analyzed in a rat model of AIONFH, and bone repair and pathological damage were evaluated by Micro-CT, RT-qPCR, HE, Masson staining, and immunohistochemistry (COL I). The results showed that M2 macrophages were successfully polarized, with an average M2-Exo particle size of 156.4 nm and a concentration of 3.2E + 12 particles/mL. The expression of miR-122 in M2 macrophages is significantly higher than that in M0 macrophages, and miR-122 mimic can increase the content of miR-122 in M2-Exo. miR-122 in M2-Exo can promote osteogenic differentiation of rat bone marrow BMSCs, enhance cell viability, and increase the expression of osteogenesis-related genes. After being applied to the AIONFH rat model, the injection of M2-exo and miR-122 mimics significantly improved the repair effect of articular cartilage, alleviated pathological changes, and promoted the regeneration of bone tissue. M2-macrophage-derived exosomal miR-122 induces osteogenic differentiation of bone mesenchymal stem cells in treating AIONFH.

Cecropin AD ameliorates pneumonia and intestinal injury in mice with mycoplasma pneumoniae by mediating gut microbiota

Animals infected with mycoplasma pneumoniae not only develop respiratory diseases, but also cause digestive diseases through the lung-gut axis mediated by the intestinal flora, and vice versa. Antimicrobial peptides are characterized by their bactericidal, anti-inflammatory, and intestinal flora-regulating properties. However, the effect of cecropin AD (CAD) against mycoplasma pneumonia remains unclear. To investigate the anti-inflammatory effect of CAD on mycoplasma pneumonia and the associated mechanism, mice were infected with Mycoplasma capricolum subsp. Capripneumoniae(Mccp) to elicit lung inflammation, followed by oral administration of CAD via gavage. The findings showed that mice receiving twice injections of 2.08 × 108 copies of Mccp suffered significant pathological damage to their lungs and colons. Additionally, there was a notable upsurge in inflammatory factors within the affected tissues. 16 S rDNA sequencing revealed alterations in the colonic microbiota, including a decrease in the abundance of beneficial bacteria such as Corynebacterium_glutamicum and Candidatus_Saccharimonas, and an increase in the abundance of potential pathogens like Lachnospiraceae_NK4A136_group and Escherichia-Shigella. As a result, there were abnormal rises in lipopolysaccharide (LPS) levels in both colonic content and blood. Moreover, CAD treatment reversed the microbial dysbiosis and decreased the LPS levels induced by Mccp, thereby suppressing the activation of the TLR-4/NF-κB pathway and the Fas/FasL-caspase-8/-3 pathway. Consequently, this significantly mitigated the morphological and functional damage to the lungs and colons caused by Mccp. The findings offer novel insights and approaches for the clinical management of Mccp infections.

VEGFR3 mitigates hypertensive nephropathy by enhancing mitophagy via regulating crotonylation of HSPA1L

Oxidative stress-associated proximal tubular cells (PTCs) damage is an important pathogenesis of hypertensive renal injury. We previously reported the protective effect of VEGFR3 in salt-sensitive hypertension. However, the specific mechanism underlying the role of VEGFR3 in kidney during the overactivation of the renin-angiotensin-aldosterone system remains unclear. In the present study, hypertensive nephropathy was established by angiotensin II (Ang II). We found that VEGFR3 was highly increased in PTCs of Ang II-infused mice. Activation of VEGFR3 mitigated renal dysfunction, pathological damage, and oxidative stress in Ang II-induced hypertensive mice. Moreover, we found that VEGFR3 restored mitophagy deficiency induced by Ang II both in vivo and in vitro to alleviate oxidative stress injury in PTCs. Furthermore, in vitro experiment demonstrated that VEGFR3 improved abnormal mitophagy by enhancing PARKIN mitochondrial translocation. LC-MS/MS and Co-IP assays identified HSPA1L as the interacted protein of VEGFR3, which promoted the mitochondrial translocation of PARKIN. Mechanistically, VEGFR3 disorder domain bound to HSPA1L, and crotonylation modification of HSPA1L at K130 by VEGFR3 was required for mitophagy regulation in the context of Ang II-induced PTCs. Finally, the protective effect of VEGFR3 on mitophagy and oxidative stress were attenuated by transfection K130 (HSPA1L-K130R) mutant plasmid in vivo and in vitro. These findings indicated that VEGFR3 alleviated oxidative stress by promoting PARKIN-dependent mitophagy pathway via regulating HSPA1L crotonylation at K130 site in Ang II-induced PTCs, which provided a mechanistic basis for the therapeutic target in hypertensive renal injury.

Lactobacillus fermentum 016 Alleviates Mice Colitis by Modulating Oxidative Stress, Gut Microbiota, and Microbial Metabolism

Ulcerative colitis (UC) is a chronic and progressive inflammatory gastrointestinal disease closely associated with gut microbiota dysbiosis and metabolic homeostasis disruption. Although targeted microbial therapies are an emerging intervention strategy for inflammatory bowel disease (IBD), the mechanisms by which specific probiotics, such as Lactobacillus fermentum 016 (LF), alleviate UC remain unclear. The current study evaluated the effects of LF supplementation on gut health in a basal model using C57BL/6 mice. Subsequently, the preventive effects and mechanisms of LF supplementation on DSS-induced UC were systematically investigated. According to our findings, LF supplementation revealed immunoregulatory capabilities with significantly altered gut the composition of microbiota and metabolic activities, particularly enhancing tryptophan metabolism. In the UC model, LF supplementation effectively mitigated weight loss, increased the disease activity index (DAI), and alleviated diarrhea, rectal bleeding, and colon shortening. Moreover, it reduced colonic pathological damage and histological injury scores. LF intervention improved antioxidant markers and intestinal mucosal barrier function with the activation of the Nrf2–Keap1 signaling pathway and regulation of systemic inflammatory markers, i.e., IL-1β, IL-6, TNF-α, IFN-γ, IL-4, and IL-10. Importantly, LF supplementation reversed metabolic disturbances by significantly increasing the abundance of beneficial genera (e.g., g_Dubosiella, g_Faecalibaculum, g_Odoribacter, g_Candidatus_saccharimonas, g_Roseburia, and g_Eubacterium_xylanophilum_group) and elevating tryptophan metabolites (e.g., melatonin, kynurenic acid, 3-indoleacetic acid, 5-methoxytryptophan, and 5-hydroxyindoleacetic acid). In conclusion, Lactobacillus fermentum 016 exhibits potential for regulating gut microbiota homeostasis, enhancing tryptophan metabolism, and alleviating UC, providing critical insights for developing probiotic-based precision therapeutic strategies for IBD.

Piplartine alleviates sepsis-induced acute kidney injury by inhibiting TSPO-mediated macrophage pyroptosis.

Sepsis-induced acute kidney injury (SI-AKI) is the most common organ dysfunction of sepsis, characterized with prolonged hospitalization periods and significantly elevated mortality rates. Piplartine (PLG), an alkaloid extracted from Piper longum within the Piperaceae family, has exhibited diverse pharmacological activities, including anti-inflammatory, anti-atherosclerotic, and anti-tumor effects. Herein, we investigated whether the PLG could reverse SI-AKI and explore its possible anti-inflammatory mechanisms. We constructed an SI-AKI model using cecal ligation and puncture (CLP) and systematically evaluated the protective effect of PLG administered by gavage in the SI-AKI mice. Subsequently, we performed proteomic sequencing of the kidney and integrated data from the GeneCards and SwissTargetPrediction databases to identify potential targets and mechanisms. Immunofluorescence and western blotting were used to examine the expression of relevant targets and pathways in vivo and in vitro. The influence of PLG on the predicted target and pathway was verified using an agonist of the target protein and a series of biochemical experiments. PLG exhibited significant efficacy against pathological damage, neutrophil and macrophage infiltration, and macrophage pyroptosis in kidneys at 30 mg/kg. An integrated analysis of proteomic data identified the translocator protein (TSPO) as a potential target for the renoprotective effects of PLG. Moreover, a TSPO agonist (RO5-4864) prominently reversed the protective effect of PLG in SI-AKI mice, as manifested by a deterioration in renal function, histopathological lesions and macrophage pyroptosis in the kidneys. Our results suggest that PLG may ameliorate SI-AKI, potentially through partial inhibition of the TSPO-macrophage pyroptosis pathway.